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Applied Microbiology and Biotechnology

, Volume 62, Issue 4, pp 356–361 | Cite as

2-(2′-Hydroxyphenyl)benzene sulfinate desulfinase from the thermophilic desulfurizing bacterium Paenibacillus sp. strain A11-2: purification and characterization

  • J. KonishiEmail author
  • K. Maruhashi
Original Paper

Abstract

2-(2′-Hydroxyphenyl)benzene sulfinate (HPBSi) desulfinase (TdsB), which catalyzes the final step of desulfurization of dibenzothiophene (DBT), was purified from a thermophilic DBT- and benzothiophene (BT)-desulfurizing bacterium: Paenibacillus sp. strain A11-2. The molecular mass of the purified enzyme was 31 kDa and 39 kDa by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively, suggesting a monomeric structure. The optimal temperature and pH for the reaction involving TdsB was 55°C and the enzyme was more resistant to heat treatment than DszB, a counterpart purified from Rhodococcus erythropolis. The optimum pH for TdsB activity was pH 8. TdsB converted HPBSi to 2-hydroxybiphenyl (2-HBP) and sulfite stoichiometrically. The Km and kcat values for HPBSi were 0.33 mM and 0.32 s−1, respectively. TdsB was inactivated by SH reagents such as p-chloromercuribenzoic acid and 5,5′-dithio-bis-2-nitrobenzoic acid, but was not inhibited by chelating reagents such as EDTA and o-phenanthroline. TdsB was also inhibited by o-hydroxystyrene, the final desulfurized product of BT. However, 2-HBP and its derivatives showed only a weak inhibitory effect. TdsB desulfurized 2-(2′-hydroxyphenyl)ethen-1-sulfinate to yield o-hydroxystyrene, but DszB could not. A site-directed mutagenesis study revealed the cysteine residue at position 17 to be essential to the catalytic activity of TdsB.

Keywords

Desulfurization Organosulfur Compound Rhodococcus Erythropolis Hydroxy Compound Biodesulfurization 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

We thank all the staff of the Bio-Refining Process Laboratory for discussions. This work was supported by the Ministry of Economy, Trade and Industry, Japan.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Bio-Refining Process LaboratoryJapan Cooperation CenterShimizuJapan

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